Experimental Study Of Bidirectional Regulation Of Dangguibuxue Decoction On Different Sates Of Vascular Endothelial Cell Proliferation And Its Mechanism

Objective:This experiment research the effect of Dangguibuxue Decoction on the hypoxic vascular endothelial cells and tumor vascular endothelial cells to explore its bi-directional regulation of vascular endothelial cell proliferation and possible mechanism, to reveal the mechanism of Dangguibuxue Decoction bi-directional regulation of different diseases on angiogenesis. and to enrich the pharmaceutical theory of bi-directional regulation provide the ideas and methods.Methods:After different doses of Dangguibuxue Decoction effecting on hypoxic vascular endothelial cells (EA. Hy926) model and vascular endothelial cells (EA. Hy926) co-cultured with the tumor cells (A549) model. CCK-8assay for the proliferation of hypoxic vascular endothelial cells and vascular endothelial cells co-cultured with the tumor cells; ELISA assay for VEGF and its receptors (VEGFR1, VEGFR2, sVEGFRl. sVEGFR2) expression of hypoxic vascular endothelial cells and vascular endothelial cells co-cultured with the tumor cells.Results:1. The effect of vascular endothelial cells proliferation by Dangguibuxue Decoction bi-directional regulation1.1Compared with normal group, the hypoxic endothelial cell proliferative capacity was significantly decreased (P<0.01); Dangguibuxue Decoction each dose group could promote the proliferation of normal endothelial cells and hypoxic endothelial cells, and positively correlated with the dose, which promote hypoxic endothelial cell proliferation rates were significantly greater than the magnitude of the promotion of normal endothelial cell proliferation, compared with model group, each dose group was significantly different (P<0.01).1.2Compared with normal group, Vascular endothelial cells co-cultured with tumor cells proliferation capacity increased (P<0.01):Dangguibuxue Decoction each dose group could promote the proliferation of normal endothelial cells, inhibit the proliferation of vascular endothelial cells co-cultured with tumor cells, and positive correlate with doses, compared with model group, each dose group are significantly different (P<0.01).2. VEGF and its receptors(VEGFR1、VEGFR2、sVEGFR1、sVEGFR2) expression2.1In the expression of VEGF. Dangguibuxue Decoction each dose can promote hypoxic vascular endothelial cells and vascular endothelial cell co-cultured with tumor cells expression of VEGF. compared with model group, the hypoxia of high-dose group differences (P<0.05):co-cultured of high dose group and dose group significant differences (P<0.01).2.2In the expression of VEGFR. compared with normal group, the hypoxia model group expression of VEGFR1、VEGFR2have a slight increase (P>0.05). while Dangguibuxue Decoction groups can significantly up-regulate VEGFR1、 VEGFR2expression, and showed a dose relationship: co-culture model group was significantly up-regulate VEGFR、VEGFR2expression, the groups Dangguibuxue Decoction inhibit the expression of VEGFR1、VEGFR2and showed a dose relationship.2.3In the expression of sVEGFR, compared with normal group, the hypoxic vascular endothelial cell model group in sVEGFR1、sVEGFR2expression are significantly increased (P<0.01), while Dangguibuxue Decoction each dose group inhibit sVEGFR1、sVEGFR2expression. Co-cultured with tumor vascular endothelial cells in model group decreased expression of sVEGFR1、sVEGFR2, each dose group of Dangguibuxue Decoction can promote sVEGFR1、sVEGFR2expression, and positively correlated with dose.Conclusion:Dangguibuxue Decoction bi-directional regulation of the different states of vascular endothelial cell proliferation, the mechanism may be combined with the regulation of VEGF and VEGFR, sVEGFR two different receptors. 1、Dangguibuxue Decoction can hypoxic vascular endothelial cell proliferation, inhibit the proliferation of co-cultured vascular endothelial cells with tumor, showing a bi-directional regulation of vascular endothelial cell proliferation in different states.2、Dangguibuxue Decoction can promote endothelial cell expression of VEGF in different states, but can be selective regulation the expression of VEGF receptor (VEGFR/sVEGFR) of different pathological conditions.Therefore the mechanism of Dangguibuxue Decoction bi-directional regulation of vascular endothelial cell proliferation may be independent of endothelial cell VEGF secretion, but the selective regulation of VEGF and its receptors.